%0 Journal Article %A Swaving, Jelto %A Weijers, Carel A. G. M. %A Ooyen, Albert J. J. van %A Bont, Jan A. M. de %T Complementation of Xanthobacter Py2 mutants defective in epoxyalkane degradation, and expression and nucleotide sequence of the complementing DNA fragment %D 1995 %J Microbiology, %V 141 %N 2 %P 477-484 %@ 1465-2080 %R https://doi.org/10.1099/13500872-141-2-477 %K complementation %K Xanthobacter Py2 %K epoxyalkane degradation %I Microbiology Society, %X SUMMARY: Three Xanthobacter Py2 mutants (M3, M8 and M10) lacking epoxyalkane-degrading activity were isolated and characterized. All mutants were able to grow on acetone, the degradation product of 1,2-epoxypropane conversions. Furthermore, they contained the unidentified ‘low molecular mass fraction’ (LMF) necessary for epoxyalkane-degrading activity. Three cosmids from a gene bank complemented the mutation in M10 and M8 but not in mutant M3. Epoxyalkane-degrading activity in crude extracts of 1,2-epoxypropane-grown complemented mutants was similar to the wild-type activity. Surprisingly, M10 transformed with complementing cosmid pEP9 showed a constitutively expressed epoxyalkane-degrading activity, which was not observed in the wild-type strain. The cosmid pEP9 was conjugated into Xanthobacter autotrophicus GJ10, which is not able to degrade 1,2-epoxypropane. In crude extracts of X . autotrophicus GJ10(pEP9), epoxyalkane-degrading activity was demonstrated, but only after the addition of the LMF from Xanthobacter Py2. Hybridization experiments demonstrated an overlap on complementing cosmids pEP1, pEP3 and pEP9. Subcloning revealed a 4.8 kb Eco RI- Hin dlll fragment to be necessary for complementing the mutant M10. In the sequence of this fragment four different ORFs were found. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/13500872-141-2-477